Pentachlorophenol impairs the antimicrobic capability of blood clam via undermining humoral immunity and disrupting humoral-cellular crosstalk.

Due to past massive usage and persistent nature, pentachlorophenol (PCP) residues are prevalent in environments, posing a potential threat to various organisms such as sessile filter-feeding bivalves. Although humoral immunity and its crosstalk with cellular one are crucial for the maintaining of robust antimicrobic capability, little is known about the impacts of PCP on these critical processes in bivalve mollusks. In this study, pathogenic bacterial challenge and plasma antimicrobic capability assays were carried out to assess the toxic effects of PCP on the immunity of a common bivalve species, blood clam (Tegillarca granosa). Moreover, the impacts of PCP-exposure on the capabilities of pathogen recognition, hemocyte recruitment, and pathogen degradation were analyzed as well. Furthermore, the activation status of downstream immune-related signalling pathways upon PCP exposure was also assessed. Data obtained illustrated that 28-day treatment with environmentally realistic levels of PCP resulted in evident declines in the survival rates of blood clam upon Vibrio challenge along with markedly weakened plasma antimicrobic capability. Additionally, the levels of lectin and peptidoglycan-recognition proteins (PGRPs) in plasma as well as the expression of pattern recognition receptors (PRRs) in hemocytes were found to be significantly inhibited by PCP-exposure. Moreover, along with the downregulation of immune-related signalling pathway, markedly fewer chemokines (interleukin 8 (IL-8), IL-17, and tumor necrosis factor α (TNF-α)) in plasma and significantly suppressed chemotactic activity of hemocytes were also observed in PCP-exposed blood clams. Furthermore, compared to that of the control, blood clams treated with PCP had markedly lower levels of antimicrobic active substances, lysozyme (LZM) and antimicrobial peptides (AMP), in their plasma. In general, the results of this study suggest that PCP exposure could significantly impair the antimicrobic capability of blood clam via undermining humoral immunity and disrupting humoral-cellular crosstalk.

Potential acetylcholine-based communication in honeybee haemocytes and its modulation by a neonicotinoid insecticide.

There is growing concern that some managed and wild insect pollinator populations are in decline, potentially threatening biodiversity and sustainable food production on a global scale. In recent years, there has been increasing evidence that sub-lethal exposure to neurotoxic, neonicotinoid pesticides can negatively affect pollinator immunocompetence and could amplify the effects of diseases, likely contributing to pollinator declines. However, a direct pathway connecting neonicotinoids and immune functions remains elusive. In this study we show that haemocytes and non-neural tissues of the honeybee Apis mellifera express the building blocks of the nicotinic acetylcholine receptors that are the target of neonicotinoids. In addition, we demonstrate that the haemocytes, which form the cellular arm of the innate immune system, actively express choline acetyltransferase, a key enzyme necessary to synthesize acetylcholine. In a last step, we show that the expression of this key enzyme is affected by field-realistic doses of clothianidin, a widely used neonicotinoid. These results support a potential mechanistic framework to explain the effects of sub-lethal doses of neonicotinoids on the immune function of pollinators.

Physiological responses of the stingless bee Partamona helleri to oral exposure to three agrochemicals: impact on antioxidant enzymes and hemocyte count.

Agrochemicals pose significant threats to the survival of bees, yet the physiological impacts of sublethal doses on stingless bees remain poorly understood. This study investigated the effects of acute oral exposure to three commercial formulations of agrochemicals [CuSO4 (leaf fertilizer), glyphosate (herbicide), and spinosad (bioinsecticide)] on antioxidant enzymes, malondialdehyde content (MDA), nitric oxide (NO) levels, and total hemocyte count (THC) in the stingless bee Partamona helleri. Foragers were exposed to lethal concentrations aimed to kill 5% (LC5) of CuSO4 (120 µg mL-1) or spinosad (0.85 µg mL-1) over a 24-h period. Glyphosate-exposed bees received the recommended label concentration (7400 µg mL-1), as they exhibited 100% survival after exposure. Ingestion of CuSO4 or glyphosate-treated diets by bees was reduced. Levels of NO and catalase (CAT) remained unaffected at 0 h or 24 h post-exposure. Superoxide dismutase (SOD) activity was higher at 0 h compared to 24 h, although insignificantly so when compared to the control. Exposure to CuSO4 reduced glutathione S-transferase (GST) activity at 0 h but increased it after 24 h, for both CuSO4 and glyphosate. MDA levels decreased after 0 h exposure to CuSO4 or spinosad but increased after 24 h exposure to all tested agrochemicals. THC showed no difference among glyphosate or spinosad compared to the control or across time. However, CuSO4 exposure significantly increased THC. These findings shed light on the physiological responses of stingless bees to agrochemicals, crucial for understanding their overall health.

Differential toxic effects of nano-titanium dioxide on clams (Meretrix meretrix) with various individuality.

Nano-TiO2 is inevitably released into aquatic environment with increasing of nanotechnology industries. Study pointed that different individuality showed divergent behavioral and physiological response when facing environmental stress. However, the effects of nano-TiO2 on tolerance of bivalves with different individualities remain unknown. In the study, clams were divided into two types of individuality - proactive and reactive by post-stress recovery method. It turned out that proactive individuals had quicker shell opening level, stronger burrowing behavior, faster feeding recovery, higher standard metabolic rate and more rapid ammonia excretion ability than reactive individuals after exposed to air. Then, the survival rate, hemocytes response and oxidase activity of classified clams were evaluated after nano-TiO2 exposure. Results showed that after 30 d exposure, proactive individuals accelerated burrowing behavior with higher survival rate. Moreover, proactive clams had better adaptability and less hemocytes response and oxidative damage than reactive clams. The study highlights the individualities of marine shell fish determine individual capacity to adapt to environmental changes, play important roles in aquaculture and coastal ecosystem health.

Dragon fruit-derived oligosaccharides alter hemocyte-mediated immunity and expression of genes related to innate immunity and oxidative stress in Daphnia magna.

Dragon fruit oligosaccharide (DFO) is an indigestible prebiotic that enhances the growth and reproduction of Daphnia magna, increases the expression of genes involved in immunity, and reduces oxidative stress. This study investigated the effects of DFO on the expression of innate immunity- (Toll, Pelle, proPO, A2M, and CTL), oxidative stress- (Mn-SOD), and nitric oxide (NO) synthesis-related genes (NOS1, NOS2, and arginase) as well as NO localization and number of hemocytes in D. magna. For this ten-day-old D. magna were treated with 0 or 9 mg l-1 of DFO for 24 and 85 h. Gene expression levels, NO intensity and localization, and total hemocytes were evaluated. After 24 h, the expression of Toll and proPO increased significantly (p < 0.05), while that of C-type lectins (CTL) was reduced (p < 0.05). At 85 h, Mn-SOD and CTL expressions were markedly suppressed (p < 0.05). NO was mostly localized in the foregut, midgut, hindgut, and carapace. The expression of NOS1 was reduced after 24 h (p < 0.05). In addition, NO intensity at 24 h was insignificantly lower than the control (p > 0.05). At 85 h, the expression of NOS1, NOS2, and arginase was higher than control, but NO intensity did not differ significantly (p > 0.05). Furthermore, the total hemocyte count elevated remarkably at 85 h (p < 0.05). Our study suggested that 9 mg l-1 of DFO could alter the expression of the genes related to innate immunity, oxidative stress, and NO synthesis in D. magna and significantly stimulate hemocyte production.

In vitro impact of Bisphenol A on the immune functions of primary cultured hemocytes of Pacific abalone (Haliotis discus hannai).

This study investigated the toxic effects of Bisphenol A (BPA) on the Pacific abalone (Haliotis discus hannai) using in vitro assays with primary cultured hemocytes. The abalone hemocytes were exposed to BPA concentrations up to 100 µM to assess cytotoxicity. Subsequently, hemocytes were exposed to sublethal BPA concentrations (LC20 = 2.3 µM and LC50 = 5.8 µM) for 48 h, and we evaluated the cellular immune responses of hemocytes via flow cytometry. Results showed no significant differences between LC20 and control groups, but LC50 exposure significantly reduced phagocytosis and oxidative capacities while increasing nitric oxide production. These findings suggest that BPA exposure negatively affects the immune system of the Pacific abalone, which makes them more susceptible to infections and other stressors in their natural environment. The study also implies that in vitro assays utilizing primary cultured abalone hemocytes may serve as effective proxies for quantifying the cytotoxic effects of chemical pollutants.

Amino-modified polystyrene nanoplastics induced multiple response of Artemia hemocytes.

Polystyrene polymers cause severe toxicity to aquatic animals. However, the process and mechanisms of innate immunity of invertebrates living at the bottom of the food chain to these pollutants remain unclear. In this study, the blood system responses of zooplankton Artemia were assessed through in vivo and in vitro exposure to amino-modified polystyrene nanoplastics (PS-NH2 NPs). The results indicated that the LC50 values of PS-NH2 NPs were 1.09 µg·mL-1 over 48 h and 0.42 µg·mL-1 over 7 d. Based on the five hemocyte subpopulations identified in Artemia, in vitro exposure assays revealed that phagocytosis was performed by plasmocytes and granulocytes with phagocytic rate of 22.64 %. TEM analysis further showed that PS-NH2 NPs caused cytoplasm vacuolization, swollen mitochondria, and lipid processing disorder. Gene expression pattern results demonstrated that Spatzle, Tollip, Hsp70, Hsp90, Casp8, API5and Pxn were significantly upregulated upon acute and chronic exposure (p < 0.05), while chronic exposure could induce significantly upregulation of ProPO (p < 0.05). Moreover, PS-NH2 NPs exposure remarkably varied the hemolymph microbiota and hemogram, particularly by increasing the proportion of adipohemocytes and phagocytes (p < 0.05). Our findings suggest that PS-NH2 NPs induce different responses in Artemia hemocyte, as primarily reflected by phagocytic processes, expression of immune and apoptosis relating genes, cell fates, hemogram and hemolymph microbiota variations. These findings support the possibility of using Artemia hemocytes as bioindicator to estimate nanoplastics pollution, thus contributing to hematological toxicity research in response to nanoplastics.

Toxic effect of mussel Mytilus galloprovincialis exposed to Ag-TiO2 and ZnTi2O4-TiO2 bicomponent nanoparticles.

Nanoparticles (NPs) are widely used in various fields, including antifouling paints for ships and industrial structures submerged in water. The potential impact of NPs on aquatic organisms, particularly their potential toxicity, is a significant concern, as their negative impact has been relatively poorly studied. In this study, we evaluated the effect of different concentrations of bimetallic Ag-TiO2 and ZnTi2O4-TiO2 NPs, which could potentially be used in antifouling coatings, on the hemocytes of the Mediterranean mussel Mytilus galloprovincialis. Hemocytes were exposed to NPs at concentrations of 0.1-1 mg/L for 1 and 2 h, and the production of reactive oxygen species (ROS), levels of DNA damage, and number of dead cells were measured. Exposure to Ag-TiO2 NPs at 1 mg/L concentration for 1 h suppressed ROS production in hemocytes and reduced the relative number of agranulocytes in cell suspensions, without inducing DNA damage or cell death. Exposure to ZnTi2O4-TiO2 NPs did not cause changes in the ratio of granulocytes to agranulocytes in suspensions, nor did it affect other functional parameters of hemocytes. However, after a 2 h exposure period, ZnTi2O4-TiO2 NPs (1 mg/L) significantly reduced the production of ROS by hemocytes. These findings suggest that Ag-TiO2 and ZnTi2O4-TiO2 NPs have low acute toxicity for marine bivalves.

Octanoic acid kills Lucilia sericata (Diptera: Calliphoridae) by affecting two major defence systems: cuticular free fatty acids and immunocompetent cells.

This work examines the insecticidal activity of octanoic acid (C8:0), a short-chain fatty acid detected in entomopathogenic fungus - Conidiobolus coronatus medium, against Lucilia sericata larvae and adults. The LD50 value was calculated as 3.04±0.26 µg/mg (3040 mg/kg) of insect body mass, which places the compound in category 5 of acute toxicity (slightly hazardous). The presented research also describes its probable mechanism, with a particular focus on changes in two main insect defense mechanisms: (1) the composition of the cuticle (GC-MS analysis) and (2) immunocompetent cells (microscopic analysis of cultured hemocytes). More precisely, octanoic acid application resulted in changes in cuticular free fatty acid (FFA) profiles in both adults and larvae; generally, treatment increased short-chain FFAs, and a decrease of middle- and long-chain FFAs. Both in vivo and in vitro applications of octanoic acid resulted in vacuolisation, disintegration, and destruction of nets formed by plasmatocytes. As the compound has also previously been found to be toxic against Galleria mellonella, it appears to have lethal potential against insects in both the Orders Diptera and Lepidoptera, indicating it may have strong entomopathogenic potential. It is worth noting that octanoic acid is approved as a food additive with well-documented insecticidal activity, and hence may be a valuable component in the design of new insecticides that are safe for both humans and the environment.

Effects of the toxic dinoflagellate, Alexandrium pacificum, on the marine diatom, Chaetoceros muelleri, and mussel (Perna canaliculus) sperm and hemocytes.

Harmful algal blooms (HABs) of Alexandrium pacificum have affected the Marlborough Sounds in New Zealand since 2010, posing a threat to green-lipped mussel (GLM, Perna canaliculus) farming. Previous studies have shown A. pacificum has negative effects GLM embryos and larvae. To further investigate these toxic mechanisms, in vitro bioassays were conducted on GLM spermatozoa, hemocytes, and the diatom, Chaetoceros muelleri. The three cell types were exposed to several treatments of A. pacificum for 2 h and responses were measured using flow cytometry and pulse amplitude-modulated fluorometry. Significant spermatozoa mortality was recorded in treatments containing A. pacificum cells or fragments, while hemocyte and C. muelleri mortality was recorded in cell-free treatments of A. pacificum which contained paralytic shellfish toxins (PSTs). Variation in sensitivity between cell types as well as the sublethal effects observed, emphasise the diverse toxic mechanisms of A. pacificum on co-occurring species in the environment.

Synthesis of alga-coated copper oxide nanoparticles with potential applications in shrimp farming.

Copper (Cu) is a crucial element that plays a vital role in facilitating proper biological activities in living organisms. In this study, copper oxide nanoparticles (CuO NPs) were synthesized using a straightforward precipitation chemical method from a copper nitrate precursor at a temperature of 85 °C. Subsequently, these NPs were coated with the aqueous extract of Sargassum angustifolium algae. The size, morphology, and coating of the NPs were analyzed through various methods, revealing dimensions of approximately 50 nm, a multidimensional shaped structure, and successful algae coating. The antibacterial activity of both coated and uncoated CuO NPs against Vibrio harveyi, a significant pathogen in Litopenaeus vannamei, was investigated. Results indicated that the minimum inhibitory concentration (MIC) for uncoated CuO NPs was 1000 µg/mL, whereas for coated CuO NPs, it was 500 µg/mL. Moreover, the antioxidant activity of the synthesized NPs was assessed. Interestingly, uncoated CuO NPs exhibited superior antioxidant activity (IC50 ≥ 16 µg/mL). The study also explored the cytotoxicity of different concentrations (10-100 µg/mL) of both coated and uncoated CuO NPs. Following 48 h of incubation, cell viability assays on shrimp hemocytes and human lymphocytes were conducted. The findings indicated that CuO NPs coated with alga extract at a concentration of 10 µg/mL increased shrimp hemocyte viability. In contrast, uncoated CuO NPs at a concentration of 25 µg/mL and higher, as well as CuO NPs at a concentration of 50 µg/mL and higher, led to a decrease in shrimp hemocyte survival. Notably, this study represents the first quantitative assessment of the toxicity of CuO NPs on shrimp cells, allowing for a comparative analysis with human cells.

Copper induces cytotoxicity in freshwater bivalve Anodonta woodiana hemocytes.

Hemocytes of freshwater bivalves are an important target model for evaluating copper (Cu) toxicity in vitro, with excess Cu causing adverse responses in these organisms. Despite this, the mechanisms underlying cytotoxicity remain poorly understood. The freshwater bivalve Anodonta woodiana, employed as a model organism in freshwater environments, was utilized in this study. Hemocytes of A. woodiana were exposed to various aqueous Cu treatments (0.001, 0.01, 0.1, 1, and 10 mg/L), and a control group (no Cu added) for 3 h to investigate the cytotoxic mechanisms of Cu. The results showed a significant increase in the production of reactive oxygen species in hemocytes of all Cu exposed groups compared to the control (p < 0.05). Remarkably, Cu treatments disrupted the cellular membrane (p < 0.05) but did not induce significant changes in the stability of the lysosomal membrane. Cu targeted the mitochondria, leading to a reduction in mitochondrial membrane potential. Additionally, all Cu treatments significantly increased the degree of DNA damage (p < 0.05). Cellular damage and a significant decline in cell viability were observed when the Cu exposure concentration reached 0.1, 1, and 10 mg/L (p < 0.05). Our study provides new insights into the cytotoxicity mechanisms triggered by Cu in hemocytes of the freshwater bivalve A. woodiana, even under environmentally relevant conditions of 0.01 mg/L exposure.

A novel route of intercellular copper transport and detoxification in oyster hemocytes.

Bivalve hemocytes are oyster immune cells composed of several cellular subtypes with different functions. Hemocytes accumulate high concentrations of copper (Cu) and exert critical roles in metal sequestration and detoxification in oysters, however the specific biochemical mechanisms that govern this have yet to be fully uncovered. Herein, we demonstrate that Cu(I) is predominately sequestered in lysosomes via the Cu transporter ATP7A in hemocytes to reduce the toxic effects of intracellular Cu(I). We also found that Cu(I) is translocated along tunneling nanotubes (TNTs) relocating from high Cu(I) cells to low Cu(I) cells, effectively reducing the burden caused by overloaded Cu(I), and that ATP7A facilitates the efflux of intracellular Cu(I) in both TNTs and hemocyte subtypes. We identify that elevated glutathione (GSH) contents and heat-shock protein (Hsp) levels, as well as the activation of the cell cycle were critical in maintaining the cellular homeostasis and function of hemocytes exposed to Cu. Cu exposure also increased the expression of membrane proteins (MYOF, RalA, RalBP1, and cadherins) and lipid transporter activity which can induce TNT formation, and activated the lysosomal signaling pathway, promoting intercellular lysosomal trafficking dependent on increased hydrolase activity and ATP-dependent activity. This study explores the intracellular and intercellular transport and detoxification of Cu in oyster hemocytes, which may help in understanding the potential toxicity and fate of metals in marine animals.

Effects of Taiwanese indigenous cinnamon (Cinnamomum osmophloeum) leaf hot-water extract on nonspecific immune responses, resistance against Vibrio parahaemolyticus, nonviable cells, and haemocyte subpopulations in white shrimp (Penaeusvannamei).

This study investigated the effects of Cinnamomum osmophloeum leaf hot-water extract (CLWE) on nonspecific immune responses and resistance to Vibrio parahaemolyticus in white shrimp (Penaeus vannamei). Firstly, a cell viability assay demonstrated that the CLWE is safe to white shrimp heamocytes in the concentration of 0-500 mg L-1. Haemocytes incubated in vitro with 10 and 50 mg L-1 of CLWE showed significantly higher response in superoxide anion production, PO activity, and phagocytic activity. In the in vivo trials, white shrimp were fed with 0, 0.5, 1, 5, and 10 g kg-1 CLWE supplemented feeds (designated as CLWE 0, CLWE 0.5, CLWE 1, CLWE 5, and CLWE 10, respectively) over a period of 28 days. In vivo experiments demonstrated that CLWE 0.5 feeding group resulted in the highest total haemocyte count, superoxide anion production, phenoloxidase activity, and phagocytic activity. Moreover, CLWE 0.5 supplemented feed significantly upregulated the clotting system, antimicrobial peptides, pattern recognition receptors, pattern recognition proteins, and antioxidant defences in white shrimp. Furthermore, the shrimp were infected with V. parahaemolyticus injections after 14 days of feeding as challenge test. Based on the challenge test result, both CLWE 0.5 and CLWE 5 demonstrated a strong resistance to V. parahaemolyticus. These two dosages effectively reduced the number of nonviable cells and activated different haemocyte subpopulations. These findings indicated that treatment with CLWE 0.5 could promote nonspecific immune responses, immune-related gene expression, and resistance to V. parahaemolyticus in white shrimp.

Acetylcholinesterase and dopamine inhibition suppress the filtration rate, burrowing behaviours, and immunological responses induced by bisphenol A in the hemocytes and gills of date mussels, Lithophaga lithophaga.

Bisphenol A (BPA), a common industrial chemical with estrogenic activity, has recently gained attention due to its well-documented negative effects on humans and other organisms in the environment. The potential immunotoxicity and neurotoxicity of BPA remain poorly understood in marine invertebrate species. Therefore, the impacts of exposure to BPA on a series of behaviours, immune responses, oxidative stress, neural biomarkers, histology, and the ultrastructure of gills were investigated in the date mussel, Lithophaga lithophaga. After 28 days of exposure to 0.25, 1, 2, and 5 µg/L BPA, hemolymphs from controls and exposed date mussels were collected, and the effects of BPA on immunological parameters were evaluated. Moreover, oxidative stress and neurochemical levels were measured in the gills of L. lithophaga. BPA reduced filtration rates and burrowing behaviour, whereas a 2 µg/L BPA resulted in an insignificant increase after 24 h. The exposure of date mussels to BPA significantly increased total hemocyte counts, a significant reduction in the diameter and phagocytosis of hemocytes, as well as gill lysozyme level. BPA increased lipid peroxidation levels and SOD activity in gills exposed to 2 and 5 µg/L BPA, but decreased GSH levels and SOD activity in 0.25 and 1 µg/L BPA-treated date mussels. Dose-dependent dynamics were observed in the inhibition of acetylcholinesterase activity and dopamine levels. Histological and scanning electron microscope examination revealed cilia erosion, necrosis, inflammation, and hyperplasia formation in the gills. Overall, our findings suggest a relationship between BPA exposure and changes in the measured immune parameters, oxidative stress, and neurochemical disturbances, which may be factored into the mechanisms underlying BPA toxicity in marine molluscs, providing a scientific foundation for marine BPA risk assessment and indicating immunosuppression in BPA-exposed date mussels.

Cellular and Genomic Instability Induced by the Herbicide Glufosinate-Ammonium: An In Vitro and In Vivo Approach.

Glufosinate-ammonium (GLA), an organophosphate herbicide, is released at high concentrations in the environment, leading to concerns over its potential genotoxic effects. However, few articles are available in the literature reporting the possible cellular and nuclear effects of this compound. We assessed, by in vitro and in vivo micronucleus assays, the genotoxicity of GLA on cultured human lymphocytes and Lymnaea stagnalis hemocytes at six concentrations: 0.010 (the established acceptable daily intake value), 0.020, 0.050, 0.100, 0.200, and 0.500 µg/mL. In human lymphocytes, our results reveal a significant and concentration-dependent increase in micronuclei frequency at concentrations from 0.100 to 0.500 µg/mL, while in L. stagnalis hemocytes, significant differences were found at 0.200 and 0.500 µg/mL. A significant reduction in the proliferation index was observed at all tested concentrations, with the only exception of 0.010 µg/mL, indicating that the exposure to GLA could lead to increased cytotoxic effects. In L. stagnalis, a significant reduction in laid eggs and body growth was also observed at all concentrations. In conclusion, we provided evidence of the genomic and cellular damage induced by GLA on both cultured human lymphocytes and a model organism's hemocytes; in addition, we also demonstrated its effects on cell proliferation and reproductive health in L. stagnalis.

Photoaging Elevated the Genotoxicity of Polystyrene Microplastics to Marine Mussel Mytilus trossulus (Gould, 1850).

Micro-sized particles of synthetic polymers (microplastics) are found in all parts of marine ecosystems. This fact requires intensive study of the degree of danger of such particles to the life activity of hydrobionts and needs additional research. It is evident that hydrobionts in the marine environment are exposed to microplastics modified by biotic and abiotic degradation. To assess the toxic potential of aging microplastic, comparative studies were conducted on the response of cytochemical and genotoxic markers in hemocytes of the mussel Mytilus trossulus (Gould, 1850) after exposure to pristine and photodegraded (UV irradiation) polystyrene microparticles (µPS). The results of cytochemical tests showed that UV-irradiated µPS strongly reduced metabolism and destabilized lysosome membranes compared to pristine µPS. Using a Comet assay, it was shown that the nuclear DNA of mussel hemocytes showed high sensitivity to exposure to both types of plastics. However, the level of DNA damage was significantly higher in mussels exposed to aging µPS. It is suggested that the mechanism of increased toxicity of photo-oxidized µPS is based on free-radical reactions induced by the UV irradiation of polymers. The risks of toxic effects will be determined by the level of physicochemical degradation of the polymer, which can significantly affect the mechanisms of toxicity.

Exposure to nanopollutants (nZnO) enhances the negative effects of hypoxia and delays recovery of the mussels' immune system.

Aquatic environments face escalating challenges from multiple stressors like hypoxia and nanoparticle exposure, with impact of these combined stressors on mussel immunity being poorly understood. We investigated the individual and combined effects of short-term and long-term hypoxia and exposure to zinc oxide nanoparticles (nZnO) on immune system of the mussels (Mytilus edulis). Hemocyte functional traits (mortality, adhesion capacity, phagocytosis, lysosomal abundance, and oxidative burst), and transcript levels of immune-related genes involved in pathogen recognition (the Toll-like receptors, the complement system components, and the adaptor proteins MyD88) were assessed. Short-term hypoxia minimally affected hemocyte parameters, while prolonged exposure led to immunosuppression, impacting hemocyte abundance, viability, phagocytosis, and defensin gene expression. Under normoxia, nZnO stimulated immune responses of mussel hemocytes. However, combined nZnO and hypoxia induced more pronounced and rapid immunosuppression than hypoxia alone, indicating a synergistic interaction. nZnO exposure hindered immune parameter recovery during post-hypoxic reoxygenation, suggesting persistent impact. Opposing trends were observed in pathogen-sensing and pathogen-elimination mechanisms, with a positive correlation between pathogen-recognition system activation and hemocyte mortality. These findings underscore a complex relationship and potential conflict between pathogen-recognition ability, immune function, and cell survival in mussel hemocytes under hypoxia and nanopollutant stress, and emphasize the importance of considering multiple stressors in assessing the vulnerability and adaptability of mussel immune system under complex environmental conditions of anthropogenically modified coastal ecosystems.

Genotoxicological and physiological effects of glyphosate and its metabolite, aminomethylphosphonic acid, on the freshwater invertebrate Lymnaea stagnalis.

Aminomethylphosphonic acid (AMPA) is the main metabolite in the degradation of glyphosate, a broad-spectrum herbicide, and it is more toxic and persistent in the environment than the glyphosate itself. Owing to their extensive use, both chemicals pose a serious risk to aquatic ecosystems. Here, we explored the genotoxicological and physiological effects of glyphosate, AMPA, and the mixed solution in the proportion 1:1 in Lymnaea stagnalis, a freshwater gastropod snail. To do this, adult individuals were exposed to increasing nominal concentrations (0.0125, 0.025, 0.050, 0.100, 0.250, 0.500 µg/mL) in all three treatments once a week for four weeks. The genotoxicological effects were estimated as genomic damage, as defined by the number of micronuclei and nuclear buds observed in hemocytes, while the physiological effects were estimated as the effects on somatic growth and egg production. Exposure to glyphosate, AMPA, and the mixed solution caused genomic damage, as measured in increased frequency of micronuclei and nuclear buds and in adverse effects on somatic growth and egg production. Our findings suggest the need for more research into the harmful and synergistic effects of glyphosate and AMPA and of pesticides and their metabolites in general.

Epigallocatechin-3-gallate attenuates the sulfamethoxazole-induced immunotoxicity and reduces SMZ residues in Procambarus clarkii.

Sulfamethoxazole (SMZ) is a commonly used antibiotic in aquaculture, and its residues in water bodies pose a significant threat to aquatic organisms in the water environment. In the present study, epigallocatechin-3-gallate (EGCG), a catecholamine, was used to mitigate the immunotoxicity caused by SMZ exposure in Procambarus clarkii. EGCG reduced the apoptosis rate, which was elevated by SMZ exposure, and increased the total hemocyte count. Simultaneously, EGCG enhanced the activities of enzymes related to antibacterial and antioxidant activities, such as superoxide dismutase (SOD), catalase (CAT), lysozyme (LZM), acid phosphatase (ACP), and GSH, which were decreased following SMZ exposure. Hepatopancreatic histology confirmed that EGCG ameliorated SMZ-induced tissue damage caused by SMZ exposure. In addition to EGCG attenuating SMZ-induced immunotoxicity in crayfish, we determined that EGCG can effectively reduce SMZ residues in crayfish exposed to SMZ. In addition, at the genetic level, the expression levels of genes related to the immune response in hemocytes were disrupted after SMZ exposure, and EGCG promoted their recovery and stimulated an increase in the expression levels of metabolism-related transcripts in hemocytes. The transcriptome analysis was conducted, and "phagosome" and "apoptosis" pathways were shown to be highlighted using Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. To the best of our knowledge, this is the first study to confirm that EGCG attenuates SMZ-induced immunotoxicity in aquatic animals and reduces SMZ residues in aquatic animals exposed to SMZ. Our study contributes to the understanding of the mechanisms by which EGCG reduces the immunotoxicity of antibiotic residues in aquatic animals.